皮秒脉冲电场对宫颈癌侵袭能力影响的实验研究  

Influence of picosecond pulse electric field to invasive ability of cervical cancer

作　　者：吴丽梅[1] 华媛媛[1] 吴雨桐[1] 姚陈果[2] 张睿哲[2] 王智亮[1] 熊正爱[1] 

机构地区：[1]重庆医科大学附属第二医院妇产科,重庆400010 [2]重庆大学输配电装备及系统安全与新技术国家重点实验室,重庆400030

出　　处：《解放军医学杂志》2015年第9期699-703,共5页Medical Journal of Chinese People's Liberation Army

基　　金：国家自然科学基金青年基金(81301928;81201745);重庆市卫生局医学科研项目(2012-2-068;2011-2-155)~~

摘　　要：目的研究不同场强皮秒脉冲电场（ps PEF）对宫颈癌侵袭能力的影响。方法 24只BALB/c裸鼠建立宫颈癌荷瘤裸鼠模型,待肿瘤最大径长至0.8～1.0cm时将其随机均分为：对照组（不予脉冲电场处理）及低场强（50k V/cm）、中场强（60k V/cm）和高场强（70k V/cm）组（n=6）,后3组分别给予场强ps PEF处理。1周后处死小鼠,行HE染色和电镜观察肿瘤的组织形态学变化,免疫组化染色检测肿瘤组织内血管内皮生长因子（VEGF）和基质金属蛋白酶-9（MMP-9）的表达,Western blotting分析VEGF和MMP-9蛋白水平的变化。结果 ps PEF处理后,HE染色观察到组织的坏死面积随脉冲电场强度的增加逐渐增大,电镜下可见随着脉冲电场强度的增加肿瘤细胞呈现出不同程度的凋亡和坏死。免疫组化检测结果显示,随着脉冲电场强度的增加,肿瘤组织内VEGF和MMP-9阳性细胞数逐渐减少,VEGF和MMP-9蛋白平均光密度值较对照组明显下降,且随着脉冲电场强度的增强,平均光密度值逐渐下降,组间差异有统计学意义（P〈0.05）。Western blotting结果显示,VEGF和MMP-9蛋白表达水平随着脉冲电场强度的增加逐渐降低,组间差异有统计学意义（P〈0.05）。结论 ps PEF具有抗宫颈癌效应,能抑制宫颈癌细胞分泌的VEGF和MMP-9,降低宫颈癌细胞的侵袭能力。Objective To investigate the influence of picosecond pulse electric field （psPEF） to the invasive ability of cervical cancer. Methods The model of cervical cancer was reproduced in BALB/c nude mice （n=24）, and they were randomly divided into four groups （n=6） when the xenografts had grown reaching a diameter of 0.8-1.0cm： control group （psPEF was not given）, low field intensity group （50kV/cm）, moderate field intensity group （60kV/cm） and high field intensity group （70kV/cm）. Seven days after the psPEF treatment, the histomorphological changes were observed with HE staining and transmission electron microscopy （TEM）, the expressions of vascular endothelial growth factor （VEGF） and matrix metalloproteinases-9 （MMP-9） were determined with immunohistochemical （IHC） staining, and the changes in protein level of VEGF and MMP-9 were assessed with Western blotting. Results After psPEF treatment, the area of necrosis was found to be increased with an increase in psPEF intensity. With TEM different degrees of apoptosis and necrosis in tumor cells with an increase of psPEF intensity were found. IHC showed that the number of VEGF and MMP-9 positive cells in cancer tissue was decreased with an increase in psPEF intensity. The average optical density （AOD） of VEGF and MMP-9 proteins decreased significantly in psPEF treatment groups compared with that in control group, and the AOD values in psPEF treatment groups decreased with an increase in psPEF intensity, and the decrease was statistically significant （P〈0.05）. Western blotting showed the expressive levels of VEGF and MMP-9 proteins declined gradually with an increase in psPEF intensity, and the difference between groups was statistically significant （P〈0.05）. Conclusion psPEF may have anti-cervical cancer effects by inhibiting the secretion of VEGF and MMP-9 and reducing the invasive ability of cervical cancer cells.

关 键 词：宫颈肿瘤 皮秒脉冲电场 基质金属蛋白酶9 血管内皮生长因子类 

分 类 号：R454.1[医药卫生—治疗学] R737.33[医药卫生—临床医学]